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[委内瑞拉苏克雷州库马纳市腹泻患者中溶组织内阿米巴和迪斯帕内阿米巴菌株的流行病学及分子研究]

[Epidemiologic and molecular study of Entamoeba histolytica and Entamoeba dispar strains in pacients with diarrhea in Cumana, Sucre state, Venezuela].

作者信息

Mora Leonor, García Ana, De Donato Marcos, Urdaneta Haideé

机构信息

Postgrado de Biología Aplicada, Universidad de Oriente. Núcleo de Sucre, Cumaná, Venezuela.

出版信息

Invest Clin. 2008 Jun;49(2):225-37.

Abstract

An epidemiological and molecular study on E. histolytica and E. dispar was carried out in 428 patients with gastrointestinal symptomatology of diarrhea from different health centers in Cumana, Sucre state. The samples were processed through: direct examination with 0.85% physiological saline solution, temporal lugol staining, trichromic staining and the Ritchie method of concentration; a sucrose gradient was used for cyst isolation. The small subunit of the 16S RNA was amplified by nested, multiplex PCR for the molecular detection. The E. histolytica/E. dispar prevalences according to the direct, Ritchie and trichromic staining methods were 20.09, 13.79 and 12.15%, respectively; while prevalences according to PCR for E. histolytica and E. dispar were 6.31% and 4.44%, respectively, also detecting four cases of mixed infection. Sequencing of the amplified fragments of E. histolytica showed 100% homology with the sequences with strains from Merida (Venezuela), USA, Brazil, Mexico and GenBank. The infections by E. histolytica and E. dispar were statistically associated with age but not with sex. The presence of mucus, blood and abdominal pain were only associated to E. histolytica infection. The moderate prevalence of E. histolytica shows the endemic status of this population and warns about the potential problem as a morbidity and mortality in Sucre state. The frequency of E. dispar in this population suggests the existence of an overestimation problem in the diagnosis of amoebiasis with its clinical and epidemiological implications, and shows the poor knowledge about the true prevalences of this protozoan. The PCR allowed for the differential identification of E. histolytica and E. dispar, as well as the presence of mixed infections, making a great tool for epidemiological amoebiasis studies.

摘要

在苏克雷州库马纳不同健康中心,对428例有腹泻等胃肠道症状的患者进行了溶组织内阿米巴和迪斯帕内阿米巴的流行病学及分子研究。样本处理方法如下:用0.85%生理盐水直接检查、临时卢戈氏染色、三色染色以及里奇浓缩法;用蔗糖梯度法分离包囊。通过巢式多重PCR扩增16S RNA的小亚基进行分子检测。根据直接检查、里奇法和三色染色法,溶组织内阿米巴/迪斯帕内阿米巴的感染率分别为20.09%、13.79%和12.15%;而根据PCR检测,溶组织内阿米巴和迪斯帕内阿米巴的感染率分别为6.31%和4.44%,还检测到4例混合感染。溶组织内阿米巴扩增片段的测序结果显示与来自梅里达(委内瑞拉)、美国、巴西、墨西哥的菌株以及基因库中的序列有100%的同源性。溶组织内阿米巴和迪斯帕内阿米巴感染在统计学上与年龄相关,但与性别无关。黏液、血液和腹痛的出现仅与溶组织内阿米巴感染有关。溶组织内阿米巴的中度感染率表明该人群的地方性流行状况,并警示了苏克雷州潜在的发病和死亡问题。该人群中迪斯帕内阿米巴的感染频率表明在阿米巴病诊断中存在高估问题及其临床和流行病学影响,也显示出对这种原生动物真实感染率的了解不足。PCR能够对溶组织内阿米巴和迪斯帕内阿米巴进行鉴别诊断,以及检测混合感染情况,是阿米巴病流行病学研究的有力工具。

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